N-benzenesulfonyl-L-proline compounds, preparation method and method for using the compounds in therapy

ABSTRACT

The present invention relates to compounds selected from the group consisting of: 
     (i) the compounds of the formula                    
      in which X is a halogen atom, A is a group 
     
       
         —NH—(CH 2 ) n —NH—CO—, —NH—CH 2 — or 
       
     
     
       
         
         
             
             
         
       
     
     Q is a group                    
     R 1  is hydrogen, halogen, C 1 -C 3  alkyl or C 1 -C 5    1 -oxoalkyl, R 2  is H or OH and n is  2, 3  or  4 ; and 
     (ii) their addition salts. 
     It further relates to the process for their preparation and to their use in therapeutics, especially for combating pathological conditions involving bradykinin.

FIELD OF THE INVENTION

The present invention relates to novel compounds derived fromN-(benzenesulfonyl)-(L)-proline, to the process for their preparationand to their use in therapeutics.

These novel compounds have an antagonistic action towards bradykinin andare useful in therapeutics, particularly for the treatment of pain andinflammation and especially for the treatment of asthma, cerebraltraumatic shock and allergic rhinitis.

PRIOR ART

It is known that one of the possible treatments for certain pathologicalconditions of a painful and/or inflammatory nature (such as asthma,rhinitis, septic shock, toothache, etc.) is to inhibit the action ofcertain hormones such as bradykinin or kallidin. These peptide hormonesare in fact involved in a large number of physiological processes, someof which are closely associated with these pathological conditions.

Although no product possessing this mode of action has yet beenmarketed, numerous studies have been undertaken in order to understandthe mode of action of kinins, particularly bradykinin and its homologs,and then create compounds capable of antagonizing the bradykininreceptors. Pharmacological Reviews, vol. 44, no. 1, pages 1-80 (1992)and Biopolymers (Peptide Science), vol. 37, pages 143-155 (1995) may bementioned among the numerous publications relating to this work.

Bradykinin is a peptide hormone consisting of 9 amino acids(Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg) and kallidin is a peptide hormone(Lys-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg) which contains an additionalamino acid (Lys) compared with bradykinin. It is known that earlierstudies made it possible to obtain peptides which interact with thebradykinin receptors: some of them, like bradycor (CP.0127 fromCortech), icatibant (HOE 140 from Hoechst) [“bradycor” and “icatibant”are international non-proprietary names (INN)] or NPC 17761 (fromScios-Nova), have an inhibitory action on the binding of bradykinin toits B2 receptor. Recent publications refer to other peptides capable ofhaving an antagonistic action on bradykinin in respect of its B₂receptor; examples of these publications which may be mentioned areWO-A-97/09347, WO-A-97/09346, U.S. Pat. No. 5,610,140, U.S. Pat. No.5,620,958, U.S. Pat. No. 5,610,142 and U.S. Pat. No. 5,597,803.Furthermore, non-peptide compounds have been proposed as antagoniststowards the binding of bradykinin to its B₂ receptor, especially inEP-A-0596406, EP-A-0622361, U.S. Pat. No. 5,578,601, FR-A-2735128,JP-A-09 040662, FR-A-2737892 and WO-A-97/11069. It is also known thatcertain compounds whose structure is more or less related to those ofthe compounds referred to in the present patent application have alreadybeen described for their possible antithrombotic properties, especiallyin the publications DE-A-3617183 and EP-A-0261539.

OBJECT OF THE INVENTION

There is a need for reducing or eliminating pain and inflammation inmammals and particularly in man.

To meet this need, a novel technical solution has been sought which iseffective in the treatment of pain, irrespective of its origin, andespecially in the treatment of pain associated with inflammatoryphenomena or traumatisms.

According to the invention, it is proposed to provide a novel technicalsolution which involves competitive binding, at the bradykinin B₂receptor, between (i) bradykinin and related or analogous hormones, and(ii) an antagonist, and utilizes compounds of the benzenesulfonamidetype which are structurally different from the known products mentionedabove and which are capable of limiting or substantially inhibiting thebinding of bradykinin and analogous hormones to said bradykinin B₂receptor.

According to this technical solution, the novel compounds bindcompetitively to the bradykinin B₂ receptor without causing the effectsof bradykinin on this receptor (these novel compounds are said to beantagonists). This results in the appearance of a state analogous tothat observed in the absence of bradykinin, namely a reduction in painand in inflammatory reactions.

In accordance with this novel technical solution, it is proposedaccording to a first aspect of the invention to provide compoundsderived from N-(benzenesulfonyl)-(L)-proline as novel industrialproducts, according to a second aspect of the invention to provide aprocess for the preparation of these compounds, and according to a thirdaspect of the invention to provide the use of these compounds,especially in therapeutics, as analgesics and/or anti-inflammatories.

SUBJECT OF THE INVENTION

In accordance with the novel technical solution of the invention, anN-(benzenesulfonyl)-L-proline compound is recommended as a novelindustrial product, said compound being selected from the groupconsisting of:

(i) the compounds of formula I:

in which:

X is a halogen atom,

A is a divalent group

—NH—(CH₂)_(n)—NH—CO—, —NH—CH₂— or

Q is a group

R₁ is a hydrogen atom, a halogen atom, a C₁-C₃, alkyl group with alinear or branched hydrocarbon chain, or a C₁-C₅ 1-oxoalkyl group,

R₂ is a hydrogen atom or an OH group, and

n is 2, 3 or 4; and

(ii) their addition salts.

According to the invention, a process for the preparation of thecompounds of formula I and their addition salts is also recommended.

The use of a substance which antagonizes a receptor of bradykinin andanalogous hormones is also recommended, wherein a bradykinin B₂ receptorantagonist selected from the compounds of formula I and their non-toxicaddition salts is used for obtaining a drug intended for use intherapeutics to combat pathological conditions involving bradykinin orits analogs, in particular to combat pain, and especially in thetreatment or prevention of pathological conditions associated withinflammatory or painful states.

DETAILED DESCRIPTION OF THE INVENTION

In general formula I of the compounds of the invention, halogen atom isunderstood as meaning a fluorine, chlorine, bromine or iodine atom, thepreferred halogen being the chlorine atom.

C₁-C₃ alkyl group with a linear or branched hydrocarbon chain isunderstood here as meaning a methyl, ethyl, propyl or 1-methylethylgroup.

C₁-C₅ 1-oxoalkyl group is understood as meaning acetyl, 1-oxopropyl,1-oxobutyl and 2-methyl-1-oxopropyl groups.

In the compound of formula I, the nitrogen heterocycle of pyrrolidinestructure comprises 1 asymmetric carbon atom. According to theinvention, this carbon has the S configuration, which corresponds to theconfiguration of L-proline.

“Addition salts” are understood as meaning the acid addition saltsobtained by reacting a compound of formula I with a mineral acid or anorganic acid. The preferred mineral acids for salifying a basic compoundof formula I are hydrochloric, hydrobromic, phosphoric and sulfuricacids. The preferred organic acids for salifying a basic compound offormula I are methanesulfonic, benzenesulfonic, maleic, fumaric, oxalic,citric, lactic and trifluoroacetic acids.

“Room temperature” is understood here as meaning a temperature of 15 to25° C. and “temperature close to room temperature” is understood here asmeaning a temperature of 0 to 40° C. and preferably of 10 to 35° C.

The general process recommended according to the invention for thepreparation of the compounds of formula I comprises, according to afirst variant A, the steps which consist in:

(1) reacting a hydroxylated heterocyclic compound of the formula

Q-O-Met

 in which:

Met is an alkali metal, especially Na or K, and

Q is a heterocyclic group selected from the structures Het 1, Het 2, Het3 and Het 4:

R₁ being a hydrogen atom or a C₁-C₃ alkyl group, with a compound offormula II:

 in which X is a halogen atom and X₁ is a halogen atom, preferably abromine atom, in an anhydrous solvent, for example dimethylformamide, ata temperature of between 0 and 50° C., for 0.5 to 10 hours, to give acompound of formula III:

 in which Q, X and R₁ are as defined above;

(2) if necessary, if Q in the compound of formula III is the group Het 1in which R₁ is a hydrogen atom:

 reacting said compound of formula III with a halogenating agent such asN-bromosuccinimide or N-chlorosuccinimide, in an appropriate solvent,especially a halogenated solvent, an ether or an alcohol, at atemperature of between about 0 and 50° C., for 0.5 to 20 hours, to givea compound of formula III′:

 in which:

R₁ is a halogen atom, preferably bromine or chlorine;

(3) hydrolyzing the ester group of the compound of formula III or III′obtained according to one of steps (1) or (2) above, especially byreaction with aqueous sodium hydroxide solution in a miscible solventsuch as methanol, at a temperature of the order of 20 to 60° C., for 1to 5 hours, to give, after acidification, a compound of formula IV:

 in which Q and X are as defined above and R₁ is a hydrogen atom, ahalogen atom or a C₁-C₃ alkyl group;

(4) reacting the resulting compound of formula IV with the salt of anamine of the formula

 in which A is a group

—NH—(CH₂)_(n)—NH—CO—, —NH—CH₂— or

in which n is 2, 3 or 4,

in an appropriate solvent, especially dichloromethane, in the presenceof activators such as, in particular, 1-hydroxy-7-azabenzotriazole(HOAT) and 1-[3-(dimethylaminopropyl)-3-ethyl]carbodiimide (EDCI)hydrochloride, at a temperature close to room temperature (10-35° C.),for 2 to 50 hours, to give a compound of formula I in which R₂ is H:

 in which A, Q, X and R₁ are as defined above and R₂, is H; and

(5) if necessary, reacting the resulting compound of formula I with anacid to give the corresponding acid addition salt;

according to a second variant B, the steps which consist in:

(1) reacting the acid compound of formula IV, obtained for example instep (3) of variant A, in which R₁ is a hydrogen atom, a chlorine atom,a C₁-C₃ alkyl group with a linear or branched hydrocarbon chain, or aC₁-C₅ 1-oxoalkyl group, with a compound of formula VI:

 in which A is a group

—NH—(CH₂)_(n)—NH—CO—, —NH—CH₂— or

in which n is 2, 3 or 4,

under conditions analogous to those recommended for carrying out step(4) of variant A above, to give a compound of formula VII:

 in which Q, R₁, X and A are as defined in the starting, materials;

(2) reacting the resulting compound of formula VII with hydroxylamine(freed from its hydrochloride by the action, in the reaction medium, ofan aprotic strong base such as triethylamine) in an appropriate solvent,especially an aprotic solvent such as dimethyl sulfoxide (DMSO), at roomtemperature (15-25° C.), for 1 to 12 hours, to give a compound offormula VIII:

 in which Q, R₁, X and A are as defined above;

(3) acetylating the resulting compound of formula VIII, at a temperatureclose to room temperature, for 1 to 8 hours, to give the compound offormula IX:

 in which Q, R₁, X and A are as defined above; and

(4) reducing the resulting compound of formula IX by catalytichydrogenation, especially in a solvent such as methanol, in the presenceof a hydrogenation catalyst such as Lindlar's catalyst, at a temperatureclose to room temperature, under a hydrogen pressure of between 10⁵ and10⁶ Pascals, to give the compound of formula I in which Q, R₁, X and Aare as defined above and R₂ is H;

according to a third variant C, the steps which consist in:

(1) reacting the compound of formula IV, obtained according to step (3)of variant A, with an amine of the formula

H₂N—(CH₂)_(n)—NH—R₃ or

 in which n is 2, 3 or 4, and

R₃ is an amino-protecting group such as the “Boc”(1,1-dimethylethoxycarbonyl) group,

under operating conditions analogous to those recommended for carryingout step (4) of variant A, to give a compound of formula X:

 in which B is

—NH—(CH₂)_(n)—NH— or

and Q, R₁ and X are as defined above;

(2) deprotecting the amine group of the resulting compound of formula Xso as to replace the group R₂ with a hydrogen atom, for example, if R₃is the Boc group, by reaction with an acid in solution in a solvent suchas ethyl acetate, at room temperature, for 4 to 30 hours, to give thecompound of formula XI:

 in which B, Q, R₁ and X are as defined above; and

(3) reacting the resulting compound of formula XI with4-(aminoiminomethyl)-benzoic acid, under conditions analogous to thosepreviously described in step (4) of variant A above, to give thecompound of formula I in which R₂ is H:

 in which:

Q, R₁ and X are as defined above, R₂ is H and A is a group

and according to a fourth variant D, the steps which consist in:

(1) reacting the acid of the formula

in which X is a halogen,

with a compound of the formula

 in which A is a group

—NH—(CH₂)_(n)—NH—CO—, —NH—CH₂— or

in which n is 2, 3 or 4,

under reaction conditions analogous to those recommended for carryingout step (4) of variant A above, to give the compound of formula XIII:

 in which A and X are as defined above;

(2) reacting the resulting compound of formula XIII with a hydroxylatedheterocyclic derivative of the general structure Q-OH, Q being selectedfrom the structures

in which R₁ is a hydrogen atom, a halogen atom, a C₁-C₃ alkyl group or aC₁-C₅ 1-oxoalkyl group,

under operating conditions analogous to those described for carrying outstep (1) of variant A above, to give the compound of formula XIV:

 in which Q, R₁, X and A are as defined in the starting materials; and

(3) then performing, on the resulting compound of formula XIV, a seriesof reactions analogous to those previously described in steps (2), (3)and (4) of variant B above, to give the compound of formula I accordingto the invention in which Q, X, R₁ and A are as defined in the startingcompounds.

The invention will be understood more clearly from the followingdescription of (i) Preparatory Examples and (ii) results ofpharmacological tests performed with compounds according to theinvention. Of course, these details as a whole do not imply a limitationbut are given by way of illustration.

In the case of compounds which have an asymmetric carbon in theirstructure, the absence of a particular indication, or the notation(R,S), means that the compounds are racemic; in the case of compoundswhich exhibit chirality, this is indicated immediately after thenumbering of the substituent carried by said asymmetric carbon; thesymbol (R) or (S) is then used in accordance with the Cahn-Ingold-Prelogrules. The nomenclature used in the Examples is that recommended byChemical Abstracts; thus, after reaction of the acid group with anamine, certain L-proline derivatives may become2(S)-pyrrolidinecarboxamide derivatives.

In the experimental section, the “Preparations” relate to theintermediates and the “Examples” relate the products according to theinvention.

The melting points (m.p.) indicated below are generally measured using aKoffler bench and are not corrected, so they represent instantaneousmelting points.

The spectral characteristics of the nuclear magnetic resonance (NMR)signals are given for the proton (¹H) or for the 13 isotope of carbon(¹³C); the chemical shift is indicated relative to the tetramethylsilanesignal and is followed, in brackets, by the shape of the signal (s forsinglet, d for doublet, t for triplet, q for quadruplet, m formultiplet, bs for broad signal) and the number of protons correspondingto the signal. By way of indication, the ¹H NMR spectra were run at 300MHz.

PREPARATION IN-[[3-[[2-Methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]-sulfonyl]-L-prolineMethyl Ester

A solution of 10 g (67.10⁻³ mol) of2-methyl-8-hydroxyimidazo[1,2-a]-pyridine in 300 ml of dimethylformamide(DMF) is prepared and 2.02 g (67.10⁻³ mol) of an 80% suspension ofsodium hydride in oil are added. The mixture is stirred at roomtemperature for 30 min and a solution of 2.91 g (67.10⁻³ mol) ofN-[[3-(bromomethyl)-2,4-dichlorophenyl]sulfonyl]-L-proline methyl esterin 100 ml of DMF is then added. After stirring at room temperature for15 hours, the reaction mixture is poured into 500 ml of water andextracted 3 times with ethyl acetate. The combined organic phases arewashed with water, dried over sodium sulfate and concentrated underreduced pressure. The resulting crude product is purified bychromatography on silica gel (silica gel 60 of particle size 15 to 40μm) using a toluene/2-propanol mixture (96/4; v/v) as the eluent to give22.5 g of the expected product in the form of a pinkish solid(yield=67%).

M.p.=149° C.; [α]_(D) ²³=19.0° (c=1.5; CHCl₃).

PREPARATION IIN-[[3-[[2-Methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]-sulfonyl]-L-proline

A solution of 12.2 g (24.10⁻³ mol) of the compound obtained according toPreparation I in 180 ml of methanol and 120 ml of water is prepared and48 ml of 1 N sodium hydroxide solution are then added. The reactionmixture is heated to 40° C. and stirred for 1.5 hours. The methanol isevaporated off under reduced pressure, the residue is cooled, 200 ml ofwater are added and the mixture is acidified to pH 2 by the slowaddition of 1 N hydrochloric acid solution, with stirring. The medium isextracted with dichloromethane and the organic phase obtained is driedover sodium sulfate and concentrated under reduced pressure to give 11.6g of the expected product in the form of pinkish white crystals.

M.p.=135° C.; [α]_(D) ²²=−50.7° (c=1.02; CH₂Cl₂).

PREPARATION III [3-[(4-Cyanobenzoyl)amino]propyl]carbamic acid1,1-dimethylethyl Ester

A solution of 10 g (57.10⁻³ mol) of (3-aminopropyl)carbamic acid1,1-dimethylethyl ester (or N-Boc-1,3-propanediamine) in 75 ml ofdichloromethane is prepared and 15.9 ml (114.10⁻³ mol) of triethylamineare added. The mixture is cooled with an ice bath and 10.38 g (60.10⁻³mol) of 4-cyanobenzoyl chloride are added gradually. The reaction mediumis allowed to warm up to room temperature, stirred for 10 hours and thenpoured into 150 ml of water. It is extracted with dichloromethane andthe organic phase obtained is washed with 1 N hydrochloric acid solutionand then with water and is finally dried over sodium sulfate andconcentrated under reduced pressure to give 16.4 g of the expectedproduct in the form of ochre crystals (yield=94%).

¹H NMR (DMSO): 1.37 (s, 9H); 1.63 (m, 2H); 2.97 (m, 2H); 3.26 (m, 2H);6.84 (t, 1H); 7.97 (s, 4H); 8.70 (t, 1H).

PREPARATION IV N-(3-Aminopropyl)-4-cyanobenzamide Hydrochloride

16.4 g (54.10⁻³ mol) of the compound obtained according to PreparationIII are dissolved in 150 ml of ethyl acetate, and 104 ml of a solutionof hydrogen chloride containing 2.6 mol/l in ethyl acetate are thenadded. The mixture is stirred at room temperature for 24 hours and thenconcentrated under reduced pressure. The resulting crude product iswashed with ethyl ether and dried under vacuum at 40° C. to give 12.15 gof the expected product in the form of an off-white solid (yield=94%).

M.p.=176-180° C.

PREPARATION V1-[[3-[[2-Methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]-sulfonyl]-N-[3-[(4-cyanobenzoyl)amino]propyl]-2(S)-pyrrolidinecarboxamide

A solution of 5.8 g (12.10⁻³ mol) of the compound obtained according toPreparation II in 200 ml of dichloromethane is prepared and 2.53 g(13.10⁻³ mol) of EDCI [1-(3-dimethylaminopropyl)-3-ethylcarbodiimide]and 1.79 g (13.10⁻³ mol) of HOAT (1-hydroxy-7-azabenzotriazole) are thenadded. The resulting mixture is stirred for 30 min at room temperatureand a solution of 2.69 g (12.10⁻³ mol) of the compound obtainedaccording to Preparation IV in a mixture of 150 ml of dichloromethaneand 2.6 ml (24.10⁻³ mol) of N-methylmorpholine is then added. Thereaction medium is stirred for 16 hours at room temperature and thenwashed successively with water, with 0.5 N hydrochloric acid solution,with sodium bicarbonate solution and finally with water again. Afterdrying over sodium sulfate, the organic phase is concentrated underreduced pressure and the crude product obtained is purified bychromatography on silica gel using a dichloromethane/methanol mixture(97/3; v/v) as the eluent to give 6.66 g of the expected product in theform of white crystals (yield=83%).

M.p.=100-102° C.; [α]_(D) ²³=−15° (c=0.99; CH₂Cl₂).

EXAMPLE 11-[[3-[[2-Methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]-sulfonyl]-N-[3-[[4-[(amino)(hydroxyimino)methyl]benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamide

A solution of 6.53 g (9.7.10⁻³ mol) of the compound obtained accordingto Preparation V in 100 ml of dimethyl sulfoxide (DMSO) is prepared and1.36 g (19.10⁻³ mol) of hydroxylamine hydrochloride and then 2.7 ml(19.10⁻³ mol) of triethylamine are added. The reaction mixture isstirred for 3 hours at room temperature, the same amounts ofhydroxylamine hydrochloride and triethylamine are then added again andthe reaction medium is stirred for 8 hours. It is poured into 400 ml ofwater, with stirring, and the precipitate obtained is filtered off andredissolved in dichloromethane. The organic phase obtained is washedwith water and then dried over sodium sulfate. After removal of thesolvent under reduced pressure, 5.37 g of the expected product areobtained in the form of white crystals (yield=78%).

M.p.=135° C.; [α]_(D) ²²=−6.9° (c=1.04; DMSO).

PREPARATION VI1-[[3-[[2-Methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]-sulfonyl]-N-[3-[[4-[(acetoxyimino)(amino)methyl]benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamide

A solution of 5.28 g (7.5.10⁻³ mol) of the compound obtained accordingto Example 1 in 150 ml of dichloromethane is prepared and 0.96 ml(10.10⁻³ mol) of acetic anhydride is added. After stirring at roomtemperature for 15 hours, the reaction medium is concentrated underreduced pressure to give 5.48 g of the expected product in the form ofbeige crystals.

M.p.=105-107° C.; [α]_(D) ²⁰=−19° (c=1.0; CH₃OH).

EXAMPLE 21-[[3-[[2-Methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]-sulfonyl]-N-[3-[[4-(aminoiminomethyl)benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamide

A solution of 5.4 g (7.2.10⁻³ mol) of the compound obtained according toPreparation VI in 150 ml of methanol is prepared and 1.08 g of Lindlar'scatalyst (containing 5% of palladium) are added. The mixture is stirredunder a hydrogen atmosphere, under a pressure of 3.10⁵ Pascals, for 5hours, at room temperature. After the catalyst has been filtered off,the solvent is removed by evaporation under reduced pressure and thecrude product is purified by chromatography on NH₂ grafted silica gel(“Lichroprep NH₂”) using a dichloromethane/methanol mixture (95/5; v/v)as the eluent to give 2.02 g of the expected product in the form ofwhite crystals (yield=41%).

M.p.=120-125° C.; [α]_(D) ²⁵=−36° (c=1.03; CH₃OH).

EXAMPLE 31-[[3-[[2-Methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]-sulfonyl]-N-[3-[[4-(aminoimiomethyl)benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamideMethanesulfonate

A solution of 1.95 g (2.8.10⁻³ mol) of the compound obtained accordingto Example 2 in 14 ml of methanol is prepared. 184 μl (2.8.10⁻³ mol) ofmethanesulfonic acid are added, with stirring. After stirring for 30 minat room temperature, the reaction medium is poured into 500 ml of ethylether. The precipitate obtained is filtered off, washed with ether anddissolved in 60 ml of distilled water. The solution obtained islyophilized to give 2.07 g of the expected product in the form of whitecrystals (yield 94%).

M.p.=170-172° C.; [α]_(D) ²⁵=−38.2° (c=1.03;CH₃OH).

PREPARATION VII 4-(4-Cyanobenzoyl)-1-piperazinecarboxylic Acid1,1-Dimethylethyl Ester

The expected product is obtained in the form of white crystals(yield=90%) by following a procedure analogous to Preparation III,starting from 1-piperazinecarboxylic acid 1,1-dimethylethyl ester.

¹H NMR: 1.46 (s, 9H); 3.38 (m, 4H); 3.51 (m, 2H); 3.73 (m, 2H); 7.50 (d,2H) 7.73 (d, 2H).

PREPARATION VIII 1-(4-Cyanobenzoyl)piperazine Hydrochloride

The expected product is obtained in the form of white crystals(yield=99%) by following a procedure analogous to Preparation IV,starting from the compound obtained according to Preparation VII.

M.p.=262-264° C.

PREPARATION IX1-[[3-[[2-Methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]-sulfonyl]-2(S)-[[4-(4-cyanobenzoyl)piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of white crystals(yield=93%) by following a procedure analogous to Preparation V,starting from the compound obtained according to Preparation VIII.

M.p.=112-115° C.; [α]_(D) ²¹=+2.8° (c=1.03; CH₂Cl₂).

EXAMPLE 41-[[3-[[2-Methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]-sulfonyl]-2(S)-[[4-[4-[(amino)(hydroxyimino)methyl]benzoyl]piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of white crystals(yield=84%) by following a procedure analogous to Example 1, startingfrom the compound obtained according to Preparation IX.

M.p.=165-167° C.; [α]_(D) ²³=+9.60 (c=1.04; DMSO).

PREPARATION X1-[[3-[[2-Methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-[4-[(acetoxyimino)(amino)methyl]benzoyl]piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of white crystals(yield=98%) by following a procedure analogous to Preparation VI,starting from the compound obtained according to Example 4.

M.p.=115-118° C.; [α]_(D) ²³=+10.2° (c=1; DMSO).

EXAMPLE 51-[[3-[[2-Methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-[4-(aminoiminomethyl)benzoyl]piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of pale yellow crystals(yield=60%) by following a procedure analogous to Example 2, startingfrom the compound obtained according to Preparation X.

M.p.=159-161° C.; [α]_(D) ²⁴=−30° (c=1.02; CH₃OH).

EXAMPLE 61-[[3-[[2-Methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-[4-(aminoiminomethyl)benzoyl]piperazin-1-yl]carbonyl]pyrrolidineMethanesulfonate

The expected product is obtained in the form of white crystals(yield=93%) by following a procedure analogous to Example 3, startingfrom the compound obtained according to Example 5.

M.p.=172-174° C.; [α]_(D) ²⁶=−19.3° (c=1.01; CH₃OH).

PREPARATION XIN-[[3-[[2,3-Dimethylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-L-prolineMethyl Ester

The expected product is obtained in the form of beige crystals(yield=50%) by following a procedure analogous to Preparation I,starting from 2,3-dimethyl-8-hydroxyimidazo[1,2-a]pyridine.

M.p.=128-130° C.; [α]_(D) ²⁹=−8.4° (c=0.97; C₂H5OH).

PREPARATION XIIN-[[3-[[2,3-Dimethylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-L-proline

The expected product is obtained in the form of off-white crystals(yield=60%) by following a procedure analogous to Preparation II,starting from the compound obtained according to Preparation XI.

M.p.=140-145° C.; [α]_(D) ²²=+15.8° (c=0.99; C₂H₅OH).

PREPARATION XIII1-[[3-[[2,3-Dimethylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[(4-cyanobenzoyl)amino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of beige crystals(yield=62%) by following a procedure analogous to Preparation V,starting from the compound obtained according to Preparation XII.

M.p.=128-130° C.; [α]_(D) ²⁶=−7° (c=0.95; CH₂Cl₂).

EXAMPLE 71-[[3-[[2,3-Dimethylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[[4-[(amino)(hydroxyimino)methyl]benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of off-white crystals(yield=74%) by following a procedure analogous to Example 1, startingfrom the compound obtained according to Preparation XIII.

M.p.=131-133° C.; [α]_(D) ²⁴=−9.5° (c=1.01; DMSO).

PREPARATION XIV1-[[3-[[2,3-Dimethylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[[4-[(acetoxyimino)(amino)methyl]benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of white crystals(yield=98%) by following a procedure analogous to Preparation VI,starting from the compound obtained according to Example 7.

M.p.=140-142° C.; [α]_(D) ²²=−7.5° (c=1.00; DMSO).

EXAMPLE 81-[[3-[[2,3-Dimethylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[[4-(iminoaminomethyl)benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of pale yellow crystals(yield=53%) by following a procedure analogous to Example 2, startingfrom the compound obtained according to Preparation XIV.

M.p.=150-152° C.; [α]_(D) ²⁵=−15.5° (c=1.1; CH₂Cl₂).

EXAMPLE 91-[[3-[[2,3-Dimethylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[[4-(aminoiminomethyl)benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamideMethanesulfonate

The expected product is obtained in the form of white crystals(yield=82%) by following a procedure analogous to Example 3, startingfrom the compound obtained according to Example 8.

M.p.=179-181° C.; [α]_(D) ²³=−39° (c=1.02; CH₃OH).

PREPARATION XV1-[[3-[[2,3-Dimethylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-(4-cyanobenzoyl)piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of yellow crystals(yield=41%) by following a procedure analogous to Preparation IX,starting from the compound obtained according to Preparation XII.

M.p.=125-128° C.; [α]_(D) ²⁶=+0.9° (c=0.85; CH₂Cl₂).

EXAMPLE 101-[[3-[[2,3-Dimethylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-[4-[(amino)(hydroxyimino)methyl]benzoyl]piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of cream-colored crystals(yield=76%) by following a procedure analogous to Example 4, startingfrom the compound obtained according to Preparation XV.

M.p.=179-181° C.; [α]_(D) ²⁶=−0.7° (c=1.01; CH₂Cl₂).

PREPARATION XVI1-[[3-[[2,3-Dimethylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-[4-[(acetoxyimino)(amino)methyl]benzoyl]piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of white crystals(yield=98%) by following a procedure analogous to Preparation X,starting, from the compound obtained according to Example 10.

M.p.=142-145° C.; [α]_(D) ²⁶=−13.5° (c=1.05; CH₂Cl₂).

EXAMPLE 111-[[3-[[2,3-Dimethylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-[4-(aminoiminomethyl)benzoyl]piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of white crystals(yield=47%) by following a procedure analogous to Example 5, startingfrom the compound obtained according to Preparation XVI.

M.p.=170-172° C.; [α]_(D) ²¹=−2.9° (c=1.00; CH₂Cl₂).

EXAMPLE 121-[[3-[[2,3-Dimethylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-[4-(aminoiminomethyl)benzoyl]piperazin-1-yl]carbonyl]pyrrolidineDihydrochloride

A solution of 736 mg (1.03.10⁻³ mol) of the compound obtained accordingto Example 11 in 25 ml of dichloromethane is prepared and 650 μl(2.6.10⁻³ mol) of a 4 N solution of hydrogen chloride in dioxane areadded. After stirring for 3 hours, the precipitate formed is separatedoff and taken up with diethyl ether to give a pale yellow solid, whichis filtered off. After drying under vacuum, the product is redissolvedin water and the solution is filtered and lyophilized to give 710 mg ofthe expected product in the form of a cream-colored flaky solid(yield=87%).

M.p.=225-229° C.; [α]_(D) ²⁶=+19.5° (c=1.07; CH₃OH).

EXAMPLE 131-[[3-[[2,3-Dimethylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[4-(aminoiminomethyl)phenylmethyl]-2(S)-pyrrolidinecarboxamide

A solution of 5 g (10.1.10⁻³ mol) of the compound obtained according toPreparation XII in 100 ml of dimethylformamide is prepared and 2.12 g(11.10⁻³ mol) of EDCI and 1.52 g (11.10⁻³ mol) of HOAT are added. Afterstirring for one hour at room temperature, this solution is added to asolution of 2.5 g (11.10⁻³ mol) of 4-(aminomethyl)benzenecarboximidamide[or 4-(aminoiminomethyl)benzylamine] dihydrochloride in 100 ml of DMFand 1.2 ml (11.10⁻³ mol) of N-methyl-morpholine. The reaction medium isstirred for 15 hours at room temperature. It is filtered and thefiltrate is poured into ethyl ether, with stirring. The precipitateformed in this way is separated off and redissolved in water.Dichloromethane is added and 2 N sodium hydroxide solution is added inorder to bring the medium to pH 13. The aqueous and organic phases areseparated and the organic phase is washed with water. After drying overmagnesium sulfate, the organic phase is concentrated under reducedpressure. The crude product obtained is purified by chromatography onNH₂ grafted silica gel using a dichloromethane/ethanol mixture (95/5;v/v) as the eluent to give 3.6 g of the expected product in the form ofa white solid (yield=57%).

M.p.=144-146° C.; [α]_(D) ²¹=−34.5° (c=1.03; CHCl₃).

EXAMPLE 141-[[3-[[2,3-Dimethylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[4-(aminoiminomethyl)phenylmethyl]-2(S)-pyrrolidinecarboxamideHydrochloride

A solution of 0.42 g (0.66.10⁻³ mol) of the compound obtained accordingto Example 13 in 100 ml of diethyl ether is prepared and 0.75 ml of a 1N solution of hydrogen chloride in diethyl ether is added, withstirring. The crystals formed are filtered off, washed with ether anddried under vacuum to give the expected product in the form of whitecrystals (yield=81%).

M.p.=210-214° C.; [α]_(D) ²²=−12° (c=1.00; C₂H₅OH).

PREPARATION XVIIN-[[3-[[3-Bromo-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-L-prolineMethyl Ester

A solution of 7 g (14.10⁻³ mol) of the compound obtained according toPreparation I in a mixture of 100 ml of dioxane and 100 ml of ethanol isprepared and 2.5 g (14.10⁻³ mol) of N-bromosuccinimide are added inportions at room temperature, with stirring. After stirring for 1 hour,the solvents are driven off under reduced pressure. The evaporationresidue is taken up with dichloromethane and the solution is washed withwater and then dried over magnesium sulfate and concentrated underreduced pressure. After purification by chromatography on silica gelusing a toluene/2-propanol mixture (97/3; v/v) as the eluent, andrecrystallization from 2-propanol, 7.3 g of the expected product(yield=90%) are obtained.

M.p.=146° C.; [α]_(D) ²³=−17° (c=1.5; CH₂Cl₂).

PREPARATION XVIIIN-[[3-[[3-Bromo-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-L-proline

The expected product is obtained in the form of a cream-colored powderysolid (yield=98%) by following a procedure analogous to Preparation II,starting from the compound obtained according to Preparation XVII.

M.p.=145° C.; [α]_(D) ²⁰=−30.5° (c=1.05; CH₂Cl₂).

PREPARATION XIX1-[[3-[[3-Bromo-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[1,1-dimethylethoxycarbonylamino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of a white solid(yield=95%) by following a procedure analogous to Preparation V,starting from the compound obtained according to Preparation XVIII and(3-aminopropyl)carbamic acid 1,1-dimethylethyl ester [orN-Boc-propanediamine].

M.p.=65° C.; [α]_(D) ²²=−43° (c=0.44; CH₃OH).

PREPARATION XX1-[[3-[[3-Bromo-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-aminopropyl]-2(S)-pyrrolidinecarboxamide

A solution of 0.6 g (0.83.10⁻³ mol) of the compound obtained accordingto Preparation XIX in 10 ml of dichloromethane is prepared and 89 mg(0.83.10⁻³ mol) of anisole and 3 ml of trifluoroacetic acid are added at0° C. The reaction mixture is then stirred for 4 hours at roomtemperatures after which the solvent is driven off under reducedpressure. The residue is taken up with 50 ml of water and the aqueousphase obtained is brought to pH 10 with 1 N sodium hydroxide solutionand then extracted with dichloromethane. The organic phase is washedwith water, dried over sodium sulfate and then concentrated underreduced pressure to give 0.45 g of the expected product in the form of awhite powdery solid.

M.p.=75° C.; [α]_(D) ²²=−72° (c=0.40; CH₃OH).

EXAMPLE 151-[[3-[[3-Bromo-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[4-(aminoiminomethyl)benzoylamino]propyl]-2(S)-pyrrolidinecarboxamide

A suspension of 80 mg (0.4.10⁻³ mol) of 4-(aminoiminomethyl)benzoic acidhydrochloride in 6 ml of DMF is prepared and 84 mg (0.44.10⁻³ mol) ofEDCI and 60 mg (0.44.10⁻³ mol) of HOAT are added. The mixture is stirredat room temperature for 10 min and 250 mg (0.40.10⁻³ mol) of thecompound obtained according to Preparation XX are then added. Thereaction mixture is stirred for 20 hours at room temperature and thenpoured into water. It is extracted with dichloromethane and theresulting organic phase is subsequently washed with saturated sodiumbicarbonate solution and then with water. It is dried over sodiumsulfate and concentrated under reduced pressure. The crude product ispurified by chromatography on NH₂ grafted silica gel using adichloromethane/methanol mixture (98/2; v/v) as the eluent to give 170mg of the expected product in the form of a white solid (yield=55%).

M.p.=170° C.; [α]_(D) ²²=−2.6° (c=0.16; CH₃OH).

EXAMPLE 161-[[3-[[3-Bromo-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[4-(aminoiminomethyl)benzoylamino]propyl]-2(S)-pyrrolidinecarboxamideHydrochloride

A solution of 153 mg (0.2.10⁻³ mol) of the compound obtained accordingto Example 15 in 8 ml of ethyl acetate and 2 ml of ethanol is preparedand 0.5 ml of a saturated solution of hydrogen chloride in diethyl etheris added. The mixture is stirred for 10 min and the solvents are thenremoved under reduced pressure. The residue is then redissolved inwater, the solution is filtered and the filtrate is lyophilized to give120 mg of the expected product in the form of a white powder(yield=75%).

M.p.=200° C.; [α]_(D) ²²=−13° (c=0.39; CH₃OH).

PREPARATION XXI1-[[3-[[3-Bromo-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-(1,1-dimethylethoxycarbonyl)piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of white crystals(yield=90%) by following a procedure analogous to Preparation XIX,starting from 1-piperazine-carboxylic acid 1-dimethylethyl ester.

M.p.=85° C.; [α]_(D) ²²=+9.6° (c=1.00; DMSO).

PREPARATION XXII1-[[3-[[3-Bromo-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of white crystals(yield=90%) by following a procedure analogous to Preparation XX,starting from the compound obtained according to Preparation XXI.

M.p.=178-180° C.; [α]_(D) ²¹=+16.5° (c=1.01; CH₃OH).

EXAMPLE 171-[[3-[[3-Bromo-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-[4-(aminoiminomethyl)benzoyl]piperazin-1-yl]carbonyl]pyrrolidineHydrochloride

The expected product is obtained in the form of pale yellow crystals(yield=40%) by following a procedure analogous to Example 15, startingfrom the compound obtained according to Preparation XXII, and afterpurification by reversed phase chromatography on silica gel (graftedsilica gel marketed under the name RP18) using anacetonitrile/water/hydrochloric acid mixture (75/25/1; v/v/v) as theeluent.

M.p.=200-204° C.; [α]_(D) ¹⁹=+21° (c=1.02; CH₃OH).

EXAMPLE 181-[[3-[[3-Bromo-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[4-(aminoiminomethyl)phenylmethyl]-2(S)-pyrrolidinecarboxamide

A solution of 1.13 g (2.10⁻³ mol) of the compound obtained according toPreparation XVIII in 10 ml of DMF is prepared and 382 mg (2.10⁻³ mol) ofEDCI and 272 mg (2.10⁻³ mol) of HOAT are added. The solution is stirredfor 20 min at room temperature and then added slowly to a mixture of 453mg (2.10⁻³ mol) of 4-(aminomethyl)benzenecarboximidamide dihydrochlorideand 202 mg (2.10⁻³ mol) of N-methylmorpholine in 10 ml of DMF. Thereaction mixture is stirred for 20 hours at room temperature and thenpoured into 150 ml of water. 1 N sodium hydroxide solution is added,with stirring, in order to bring the medium to pH 13, and the medium isthen extracted with dichloromethane. The organic phase is washed andthen dried over magnesium sulfate and concentrated under reducedpressure. The product obtained is purified by chromatography on silicagel (NH₂ grafted silica gel) using a dichloromethane/methanol mixture(95/5; v/v) as the eluent to give 280 mg of the expected product in theform of a white solid (yield=24%).

M.p.=124° C.; [α]_(D) ²⁵=−30° (c=0.57; CH₃OH).

EXAMPLE 191-[[3-[[3-Bromo-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[4-(aminoiminomethyl)phenylmethyl]-2(S)-pyrrolidinecarboxamideHydrochloride

The expected product is obtained in the form of a white solid(yield=92%) by following a procedure analogous to Example 16, startingfrom the compound obtained according to Example 18 (yield=92%).

M.p.=214° C.; [α]_(D) ²⁵=−27° (c=0.68; CH₃OH).

PREPARATION XXIIIN-[[3-[(2-Methylquinoxalin-8-yl)oxymethyl]-2,4-dichlorolphenyl]sulfonyl]-L-prolineMethyl Ester

The expected product is obtained in the form of white crystals(yield=92%) by following a procedure analogous to Preparation I,starting from 8-hydroxy-2-methylquinoxaline.

M.p.=158-159° C.; [α]_(D) ²⁴=−24.5° (c=1.00; CHCl₃).

PREPARATION XXIVN-[[3-[(2-Methylquinoxalin-8-yl)oxymethly]-2,4-dichlorophenyl]sulfonyl]-L-proline

The expected product is obtained in the form of off-white crystals(yield=90%) by following a procedure analogous to Preparation II,starting from the product obtained according to Preparation XXIII.

M.p.=238-240° C.; [α]_(D) ²⁶=−111° (c=0.97; CHCl₃).

PREPARATION XXV1-[[3-[(2-Methylquinoxalin-8-yl)oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[(4-cyanobenzoyl)amino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of white crystals(yield=79%) by following a procedure analogous to Preparation V,starting from the product obtained according to Preparation XXIV.

M.p.=104-106° C.; [α]_(D) ²⁶=−32° (c=0.96; CHCl₃).

EXAMPLE 201-[[3-[(2-Methylquinoxalin-8-yl)oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[[4-[(amino)(hydroxyimino)methyl]benzoyl]amino]propyl]-2-(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of white crystals(yield=88%) by following a procedure analogous to Example 1, startingfrom the product obtained according to Preparation XXV.

M.p.=172-176° C.; [α]_(D) ²³=−7.5° (c=0.98; DMSO).

PREPARATION XXVI1-[[3-[(2-Methylquinoxalin-8-yl)oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[[4-[(acetoxyimino)(amino)methyl]benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of pinkish crystals(yield=95%) by following a procedure analogous to Preparation VI,starting from the compound obtained according to Example 20.

M.p.=151-154° C.; [α]_(D) ²⁴=−10° (c=0.95; DMSO).

EXAMPLE 211-[[3-[(2-Methylquinoxalin-8-yl)oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[[4-(aminoiminomethyl)benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of pinkish crystals(yield=55%) by following, a procedure analogous to Example 2, starting,from the compound obtained according to Preparation XXVI.

M.p.=136-140° C.; [α]_(D) ²⁴=−10° (c=1.00; DMSO).

EXAMPLE 221-[[3-[(2-Methylquinoxalin-8-yl)oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[[4-(aminoiminomethyl)benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamideMethanesulfonate

The expected product is obtained in the form of light yellow crystals(yield=84%) by following a procedure analogous to Example 3, startingfrom the compound obtained according to Example 21.

M.p.=165-167° C.; [α]_(D) ²⁵=−38.2° (c=1.03; CH₃OH).

PREPARATION XXVII1-[[3-[(2-Methylquinoxalin-8-yl)oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2-(S)-[[4-(4-cyanobenzoyl)piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form white crystals (yield=96%)by following a procedure analogous to Preparation IX, starting from thecompound obtained according to Preparation XXIV.

M.p.=126-130° C.; [α]_(D) ²⁶=−0.4° (c=1.13; CHCl₃).

EXAMPLE 231-[[3-[(2-Methylquinoxalin-8-yl)oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2-(S)-[[4-[4-[(amino)(hydroxyimino)methyl]benzoyl]piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of white crystals(yield=86%) by following a procedure analogous to Example 4, startingfrom the compound obtained according to Preparation XXVII.

M.p.=210-212° C.; [α]_(D) ²⁴=+19.5° (c=0.47; DMSO).

PREPARATION XXVIII1-[[3-[(2-Methylquinoxalin-8-yl)oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-[4-[(acetoxyimino)(amino)methyl]benzoyl]piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of yellow crystals(yield=98%) by following a procedure analogous to Preparation X,starting from the compound obtained according to Example 23.

M.p.=161-165° C.; [α]_(D) ²³=+9° (c=1.02; DMSO).

EXAMPLE 241-[[3-[(2-Methylquinoxalin-8-yl)oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-[4-(aminoiminomethyl)benzoyl]piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of beige crystals(yield=45%) by following a procedure analogous to Example 5, startingfrom the compound obtained according to Preparation XXVIII.

M.p.=155-158° C.; [α]_(D) ²⁴=+9.7° (c=0.95; DMSO).

EXAMPLE 251-[[3-[(2-Methylquinoxalin-8-yl)oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-[4-(aminoiminomethyl)benzoyl]piperazin-1-yl]carbonyl]pyrrolidineHydrochloride

The expected product is obtained in the form of a white solid(yield=95%) by following a procedure analogous to Example 16, startingfrom the compound obtained according to Example 24.

M.p.=192-195° C.; [α]_(D) ²⁴=+6.7° (c=1.03, DMSO).

EXAMPLE 261-[[3-[(2-Methylquinoxalin-8-yl)oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[4-(aminoiminomethyl)phenylmethyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of a beige solid(yield=27%) by following a procedure analogous to Example 13, startingfrom the compound obtained according to Preparation XXIV.

M.p.=120-125° C.; [α]_(D) ₂₃=−42° (c=1.00; CHCl₃).

EXAMPLE 271-[[3-[(2-Methylquinoxalin-8-yl)oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[4-(aminoiminomethyl)phenylmethyl]-2(S)-pyrrolidinecarboxamideHydrochloride

A solution of 387 mg (0.62.10⁻³ mol) of the compound obtained accordingto Example 26 in 100 ml of ethyl ether is prepared and 0.7 ml of a 1 Nsolution of hydrogen chloride in ethyl ether is added. The crystalsformed are filtered off, washed with ether, dried under vacuum andredissolved in 10 ml of water. After filtration and lyophilization ofthe filtrate, 376 mg of the expected product are obtained in the form ofa white powder (yield=92%).

M.p.=180-184° C.; [α]_(D) ²³=−43° (c=0.99; ethanol).

PREPARATION XXIXN-[[3-[[2-Methyl-4H-pyrido[1,2-a]pyrimidin-9-yl-4-one]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-(L)-prolineMethyl Ester

The expected product is obtained in the forin of off-white crystals(yield=56%) by following a procedure analogous to Preparation I,starting from 9-hydroxy-2-methyl-4H-pyrido[1,2-a]pyrimidin-4-one.

M.p.=166° C.; [α]_(D) ²²=−24° (c=0.37; CHCl₃).

PREPARATION XXXN-[[3-[[2-Methyl-4H-pyrido[1,2-a]pyrimidin-9-yl-4-one]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-(L)-proline

The expected product is obtained in the form of a beige solid(yield=92%) by following a procedure analogous to Preparation II,starting from the compound obtained according to Preparation XXIX.

M.p.=150° C.; [α]_(D) ²²=−86° (c=0.4; CHCl₃).

PREPARATION XXXI1-[[3-[[2-Methyl-4H-pyrido[1,2-a]pyrimidin-9-yl-4-one]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[(4-cyanobenzoyl)amino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of a white solid(yield=83%) by following a procedure analogous to Preparation V,starting from the compound obtained according to Preparation XXX.

M.p.=112° C.; [α]_(D) ²²=−50° (c=0.31; CHCl₃).

EXAMPLE 281-[[3-[[2-Methyl-4H-pyrido[1,2-a]pyrimidin-9-yl-4-one]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-3-[4-[(amino)(hydroxyimino)methyl]benzoylamino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of a white solid(yield=87%) by following a procedure analogous to Example 1, startingfrom the compound obtained according to Preparation XXXI.

M.p.=251° C.; [α]_(D) ²²=−11° (c=0.36; DMSO).

PREPARATION XXXII1-[[3-[[2-Methyl-4H-pyrido[1,2-a]pyrimidin-9-yl-4-one]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[4-](acetoxyimino)(amino)methyl]benzoylamino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of a creamy white powder(yield=95%) by following a procedure analogous to Preparation VI,starting from the compound obtained according to Example 28.

M.p.=150° C.; [α]_(D) ²²=−24.5° (c=0.33; CHCl₃).

EXAMPLE 291-[[3-[[2-Methyl-4H-pyrido[1,2-a]pyrimidin-9-yl-4-one]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[[4-(aminoiminomethyl)benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of a white powdery solid(yield=62%) by following a procedure analogous to Example 2, startingfrom the compound obtained according to Preparation XXXII.

M.p.=153° C.; [α]_(D) ²²=−19° (c=0.32; DMSO).

EXAMPLE 301-[[3-[[2-Methyl-4H-pyrido[1,2-a]pyrimidin-9-yl-4-one]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[[4-(aminoiminomethyl)benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamideBis(methanesulfonate)

A solution of 160 mg (0.224.10⁻³ mol) of the compound obtained accordingto Example 29 in 3 ml of dichloromethane and 3 ml of ethanol isprepared. 43 mg (0.45.10⁻³ mol) of methanesulfonic acid are added andthe mixture is stirred for 10 min. 20 ml of diethyl ether are then addedand, after 5 min, the crystals formed are filtered off and washed on thefilter with ether. After drying under vacuum, the crystals areredissolved in 20 ml of water and the solution is lyophilized to give170 mg of the expected product in the form of fine white crystals(yield=85%).

M.p.=183° C.; [α]_(D) ²⁴=−32° (c=0.46; CH₃OH).

PREPARATION XXXIII 1-[4-Cyanobenzoyl]piperazine Trifluoroacetate

A solution of 40.7 g (129.10⁻³ mol) of the compound obtained accordingto Preparation VII in 500 ml of dichloromethane is prepared and 150 mlof trifluoroacetic acid are added gradually. The mixture is subsequentlystirred at room temperature for 30 min and then concentrated underreduced pressure. The residue is taken up with 400 ml of diethyl ether,with stirring; the crystals formed are filtered off and then dried undervacuum to give 30.8 g of the expected product in the form of whitecrystals (yield=87%).

M.p.=180° C.;

PREPARATION XXXIV1-(1,1-Dimethylethoxycarbonyl)-2(S)-[[4-(4-cyanobenzoyl)piperazin-1-yl]carbonyl]pyrrolidine

A solution of 10.78 g (50.10⁻3 mol) ofN-(1,1-dimethylethoxycarbonyl)-L-proline (or N-Boc-L-proline) in 90 mlof DMF is prepared and 11.47 g (60.10⁻³ mol) of EDCI and 8.18 g (60.10⁻³mol) of HOAT are added. The mixture is stirred for 30 min at roomtemperature and a solution of 15 g (55.10⁻³ mol) of the compoundobtained according to Preparation XXXIII in 90 ml of DMF and 6.08 g(60.10⁻³ mol) of triethylamine is then added. After stirring for 3 hoursat room temperature, the reaction medium is poured into iced water andextracted with dichloromethane. The organic phase is washed and thendried over magnesium sulfate and concentrated under reduced pressure.The crude product is purified by chromatography on silica gel using adichloromethane/methanol mixture (98/2; v/v) as the eluent to give 20 gof the expected product in the form of a white solid (yield=97%).

M.p.=50° C.; [α]_(D) ²³=+3° (c=0.50; CHCl₃).

PREPARATION XXXV2(S)-[4-(4-Cyanobenzoyl)piperazin-1-ylcarbonyl]pyrrolidineTrifluoroacetate

The expected product is obtained in the form of a white solid(yield=81%) by following a procedure analogous to Preparation XX,starting from the compound obtained according to Preparation XXXIV.

M.p.=50° C.; [α]_(D) ²²=−35° (c=0.58; CHCl₃).

PREPARATION XXXVI1-[(3-Bromomethyl-2,4-dichlorophenyl)sulfonyl]-2(S)-[[4-(4-cyanobenzoyl)-piperazin-1-yl]carbonyl]pyrrolidine

A solution of 13.4 g (39.5.10⁻³ mol) of3-bromomethyl-2,4-dichlorobenzenesulfonyl chloride in 35 ml ofacetonitrile is prepared and 16.9 g (39.5.10⁻³ mol) of the compoundobtained according to Preparation XXXV are added at room temperature. Asolution of 10 g (0.1 mol) of potassium bicarbonate in 40 ml of water isthen added dropwise. The reaction mixture is stirred for 20 hours, wateris then added and the mixture is extracted with ethyl acetate. Theorganic phase obtained is washed with water, dried over magnesiumsulfate and concentrated under reduced pressure. The crude product ispurified by chromatography on silica gel using a dichloromethane/ethylacetate mixture (9/1; v/v) as the eluent to give 18.8 g of the expectedproduct (partly containing its chlorinated analog) in the form of abeige solid.

M.p.=115° C.;

PREPARATION XXXVII1-[[3-[[2-Methylpyrido[1,2-a]pyrimidin-9-yl-4-one]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-(4-cyanobenzoyl)piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of white crystals(yield=62%) by following a procedure analogous to Preparation I,starting from 9-hydroxy-2-methylpyrido[1,2-a]pyrimidin-4-one and thecompound obtained according to Preparation XXXVI, and afterrecrystallization from an ethyl acetate/diisopropyl ether mixture.

M.p.=160° C.; [α]_(D) ²²=+9° (c=0.39; CHCl₃).

EXAMPLE 311-[[3-[[2-Methylpyrido[1,2-a]pyrimidin-9-yl-4-one]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-[4-[(amino)(hydroxyimino)methyl]benzoyl]piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of a white solid(yield=77%) by following a procedure analogous to Example 1, startingfrom the compound obtained according to Preparation XXXVII.

M.p.=180° C.; [α]_(D) ²²=+26° (c=0.36; CHCl₃).

PREPARATION XXXVIII1-[[3-[[2-Methylpyrido[1,2-a]pyrimidin-9-yl-4-one]oxymethyl]-2,4-dichlolophenyl]sulfonyl]-2(S)-[[4-[4-[(acetoxyimino)(amino)methyl]benzoyl]piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of a white solid(yield=93%) by following a procedure analogous to Preparation VI,starting from the compound obtained according to Example 31.

M.p.=160° C.; [α]_(D) ²²=+6° (c=0.37; DMSO).

EXAMPLE 321-[[3-[[2-Methylpyrido[1,2-a]pyrimidin-9-yl-4-one]oxymethyl]-2,4-dichlorophenyl]sulfonyl]2(S)-[[4-[4-(aminoiminomethyl)benzoyl]piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of a yellow solid(yield=44%) by following a procedure analogous to Example 2, startingfrom the compound obtained according to Preparation XXXVIII.

M.p.=168° C.; [α]_(D) ²²=+32° (c=0.38; CHCl₃).

EXAMPLE 331-[[3-[[2-Methylpyrido[1,2-a]pyrimidin-9-yl-4-one]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-[4-(aminoiminoethyl)benzoyl]piperazin-1-yl-carbonyl]pyrrolidineBis(methanesulfonate)

The expected product is obtained in the form of a white solid(yield=80%) by following a procedure analogous to Example 30, startingfrom the compound obtained according to Example 32.

M.p.=181° C.; [α]_(D) ²⁴=+13° (c=0.35; CH₃OH).

PREPARATION XXXIXN-[[3-[(2,4-Dimethylquinazolin-8-yl)oxymethyl]-2,4-dichlorophenyl]sulfonyl]-L-prolineMethyl Ester

The expected product is obtained in the form of a white solid(yield=70%) by following a procedure analogous to Preparation I,starting from 2,4-dimethyl-8-hydroxyquinazoline.

M.p.=140-142° C.; [α]_(D) ²⁴=−29° (c=1; CHCl₃).

PREPARATION XLN-[[3-[(2,4-Dimethylquinazolin-8-yl)oxymethyl]-2,4-dichlorophenyl]sulfonyl]-L-proline

The expected product is obtained in the form of a beige solid(yield=98%) by following a procedure analogous to Preparation II,starting from the compound obtained in Preparation XXXIX.

M.p.=94-96° C.;

EXAMPLE 341-[[3-[(2,4-Dimethylquinazolin-8-yl)oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[4-(aminoimiomethyl)phenylmethyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of a white powder(yield=25%) by following a procedure analogous to Example 13, startingfrom the compound obtained in Preparation XL.

M.p.=139-142° C.;

EXAMPLE 351-[[3-[(2,4-Dimethylquinazolin-8-yl)oxymethyl-2,4-dichlorophenyl]sulfonyl]-N-[4-(aminoiminomethyl)phenylmethyl]2(S)-pyrrolidinecarboxamideHydrochloride

The expected product is obtained in the form of a white powder(yield=95%) by following a procedure analogous to Example 16, startingfrom the compound obtained according to Example 34.

M.p.=177-179° C.; [α]_(D) ²¹=−43° (c=0.80; CH₃OH).

PREPARATION XLIN-[[3-[[3-Chloro-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-L-prolineMethyl Ester

The expected product is obtained in the form of a beige solid(yield=95%) by following a procedure analogous to Preparation XVII, theN-bromosuccinimide being replaced with N-chlorosuccinimide.

M.p.=60° C.; [α]_(D) ²²=+13° (c=0.35; DMSO).

PREPARATION XLIIN-[[3-[[3-Chloro-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-L-proline

The expected product is obtained in the form of a white solid(yield=88%) by following a procedure analogous to Preparation XVIII,starting from the compound obtained according to Preparation XLI.

M.p.=144° C.; [α]_(D) ²²=−60.5° (c=0.35; CHCl₃).

PREPARATION XLIII1-[[3-[[3-Chloro-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[(4-cyanobenzoyl)amino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of an off-white solid(yield=51%) by following a procedure analogous to Preparation V,starting from the compound obtained in Preparation XLII.

M.p.=115° C.; [α]_(D) ²²=−32° (c=0.60; CHCl₃).

EXAMPLE 361-[[3-[[3-Chloro-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[[4-[(amino)(hydroxyimino)methyl]benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of a white solid(yield=96%) by following a procedure analogous to Example 1, startingfrom the compound obtained in Preparation XLIII.

M.p.=155° C.; [α]_(D) ²²=−41° (c=0.40; CHCl₃).

PREPARATION XLIV1-[[3-[[3-Chloro-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[[4-[(acetoxyimino)(amino)methyl]benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of a crude solid(yield=82%) by following a procedure analogous to Preparation VI,starting from the compound obtained according to Example 36.

M.p.=170° C.; [α]_(D) ²¹=−32° (c=0.35; CHCl₃).

EXAMPLE 371-[[3-[[3-Chloro-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[[4-(aminoiminomethyl)benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of a beige solid(yield=47%) by following a procedure analogous to Example 2, startingfrom the compound obtained in Preparation XLIV.

M.p.=115° C.; [α]_(D) ²²=−9.5° (c=0.55; DMSO).

EXAMPLE 381-[[3-[[3-Chloro-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[[4-(aminoimiomethyl)benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamideBis(methanesulfonate)

The expected product is obtained in the form of a white solid(yield=90%) by following a procedure analogous to Example 30, startingfrom the compound obtained according to Example 37.

M.p.=154° C.; [α]_(D) ²⁷=−19° (c=0.32; CH₃OH).

PREPARATION XLV1-[[3-[[3-Chloro-2-methylidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-(4-cyanobenzoyl)piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of a creamy white solid(yield=49%) by following a procedure analogous to Preparation V,starting from the compounds obtained according to Preparations XLII andVIII.

M.p.=100° C.; [α]_(D) ²⁸ =−11° (c=0.35; CH₃OH).

EXAMPLE 391-[[3-[[3-Chloro-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-[4-[(amino)(hydroxyimino)methyl]benzoyl]piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of a white solid(yield=96%) by following a procedure analogous to Example 1, startingfrom the compound obtained according to Preparation XLV.

M.p.=168° C.; [α]_(D) ²⁸=−25° (c=0.59; CH₃OH).

PREPARATION XLVI1-[[3-[[3-Chloro-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-[4-[(acetoxyimino)(amino)methyl]benzoyl]piperazin-1-yl]carbonyl]pyrolidine

The expected product is obtained in the form of an off-white solid(yield=88%) by following a procedure analogous to Preparation VI,starting from the compound obtained according to Example 39.

M.p.=150° C.; [α]_(D) ²⁶=−16.5° (c=0.40; CH₃OH).

EXAMPLE 401-[[3-[[3-Chloro-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-[4-[(aminoiminomethyl)benzoyl]piperazin-1-yl]carbonyl]pyrrolidine

The expected product is obtained in the form of a pale yellow solid(yield=96%) by following a procedure analogous to Example 2, startingfrom the compound obtained according to Preparation XLVI.

M.p.=164° C.; [α]_(D) ²⁶=−14° (c=0.33; CH₃OH).

EXAMPLE 411-[[3-[[3-Chloro-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-2(S)-[[4-[4-[(aminoiminomethyl)benzoyl]piperazin-1-yl]carbonyl]pyrrolidineBis(methanesulfonate)

The expected product is obtained in the form of a fine white solid(yield=83%) by following a procedure analogous to Example 3 (in thiscase dichloromethane is used to dissolve the starting compound),starting from the compound obtained according to Example 40.

M.p.=192° C.; [α]_(D) ²⁷=+21° (c=0.42; CH₃OH).

PREPARATION XLVIIN-[[3-[(2-Aminopyridin-3-yl)oxymethyl]-2,4-dichlorophenyl]sulfonyl]-(L)-prolineMethyl Ester

A solution of 4 g (36.4.10⁻³ mol) of 2-amino-3-hydroxypyridine in 200 mlof dimethylformamide is prepared and 1.09 g (36.4.10⁻³ mol) of sodiumhydride are added at room temperature. After stirring for 30 min. 15.70, (36.4.10⁻³ mol) ofN-[(3-bromomethyl-2,4-dichlorophenyl)sulfonyl]-(L)-proline methyl esterare added and the mixture is stirred for 2 hours at room temperature.The reaction medium is then poured into 250 ml of iced water. Theproduct is filtered off and redissolved in ethyl acetate. The solutionis washed with water and then dried over sodium sulfate and concentratedunder reduced pressure. The crude product obtained is purified bychromatography on silica gel using a dichloromethane/methanol mixture(97/3; v/v) as the eluent to give 12.4 g of the expected compound in theform of orange-yellow crystals (yield=74.5%).

M.p.=68° C.; [α]_(D) ²⁵=−12.8° (c=1.05; CH₃OH).

PREPARATION XLVIIIN-[[3-[[3-Acetyl-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]prolineMethyl Ester

A solution of 10.1 g (22.10⁻³ mol) of the compound obtained according toPreparation XLVII in 160 ml of ethanol is prepared. 6 ml (50.10⁻³ mol)of 3-chloro-2,4-pentanedione are added, with stirring, and the reactionmixture is stirred at the reflux point of the solvent for 15 hours. Itis then concentrated under reduced pressure and the crude productobtained is purified by chromatography on silica gel using amethylcyclohexane/ethyl acetate mixture (1/1; v/v) as the eluent to give2.67 g of the expected product in the form of beige crystals(yield=22.5%).

M.p.=178° C.; [α]_(D) ²⁵=−8.6° (c=1.05; DMSO).

PREPARATION ILN-[[3-[[3-Acetyl-2-methylimidazol[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]proline

The expected product is obtained in the form of creamy white crystals(yield=91%) by following a procedure analogous to Preparation II,starting from the compound obtained according to Preparation XLVIII.

M.p.=162-164° C.; [α]_(D) ²⁵=−8.3° (c=1.01; CH₃OH).

PREPARATION L1-[[3-[[3-Acetyl-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[(4-cyanobenzoyl)amino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of off-white crystals(yield=94%) by following a procedure analogous to Preparation V,starting from the compound obtained in Preparation IL.

M.p.=100-102° C.; [α]_(D) ²⁰=−30.6° (c=1.02; CHCl₃).

EXAMPLE 421-[[3-[[3-Acetyl-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[[4-[(amino)(hydroxyimino)methyl]benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of white crystals(yield=88%) by following a procedure analogous to Example 1, startingfrom the compound obtained in Preparation L.

M.p.=164-166° C.; [α]_(D) ²⁵=−22° (c=1.00; CH₃OH).

PREPARATION LI1-[[3-[[3-Acetyl-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[[4-[(acetoxyimino)(amino)methyl]benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of white crystals(yield=97%) by following a procedure analogous to Preparation VI,starting from the compound obtained according to Example 42.

M.p.=115-117° C.; [α]_(D) ²⁴=−6.4° (c=1.06; DMSO).

EXAMPLE 431-[[3-[[3-Acetyl-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[[4-(aminoiminomethyl)benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamide

The expected product is obtained in the form of white crystals(yield=69%) by following a procedure analogous to Example 2, startingfrom the compound obtained in Preparation LI.

M.p.=158-160° C.; [α]_(D) ²¹=−9.5° (c=1.01; DMSO).

EXAMPLE 441-[[3-[[3-Acetyl-2-methylimidazo[1,2-a]pyridin-8-yl]oxymethyl]-2,4-dichlorophenyl]sulfonyl]-N-[3-[[4-(aminoiminomethyl)benzoyl]amino]propyl]-2(S)-pyrrolidinecarboxamideBis(methanestilfonate)

The expected product is obtained in the form of white crystals(yield=84%) by following a procedure analogous to Example 3, startingfrom the compound obtained according to Example 43.

M.p.=170-172° C.; [α]_(D) ²²=−7.5° (c=1.02; DMSO).

The activity of the products according to the invention was evaluated inrespect of their ability to bind to the bradykinin receptors. Kinins, ofwhich bradykinin is the main representative, actually form a group ofsmall peptides which make an important contribution to the inflammatoryresponse and therefore appear to be involved in the pathophlysiology ofinflammatory diseases. Furthermore, bradykinin is one of the most potentanalgesics known. Kinins activate two types of receptor, called B₁ andB₂. The B₂ receptor belongs to the large family of receptors with seventransmembrane domains coupled to the G proteins. In the presentinvention we describe compounds which bind to the B₂ receptor andthereby block the binding of bradykinin.

The following pharmacological test is used: Ileum segments are isolatedfrom male guinea-pigs [of the Dunkin-Hartley strain (Iffa Credo,l'Arbresle, France)] and ground in the following TES buffer: TES 25 mM,1,10-phenanthroline 1 mM (pH 6.8), bacitracin 140 μg/ml, BSA 1 g/l. Themembranes are then isolated by centrifugation (18,000 rpm; 20 min 4°C.). The binding studies are carried out in this TES buffer using[³H]-bradykinin (120 pM) and 50 μg of membrane protein per test (finalvolume 500 μl) with an equilibrium time of 90 min at 20° C. Thepercentage inhibition of the binding of [³H]-bradykinin is thendetermined in the presence of one of the test compounds according to theinvention at a concentration of 10⁻⁶ M.

The results obtained from these tests (shown as “activity”) are collatedin Table I below with reference to the Examples given in thedescription. In this Table, the meaning of the heterocyclic radicals Qis given in the notes and the nature of the salts is indicated by theabbreviations Chl in the case of a hydrochloric acid salt and Ms in thecase of a methanesulfonic acid salt.

The compounds according to the invention have an antagonistic effecttowards the bradykinin B₂ receptor (they inhibit the activation of theB₂ receptors which is induced by bradykinin). This property has beendemonstrated experimentally by means of pharmacological tests describedin the publication by D. PRUNEAU et al. (British Journal ofPharmacology, October 1995, vol. 116 (no. 3), pp. 2106-2112) andperformed on guilnea-pig ileum. The experimental results, evaluated bythe pK_(B) described in the above publication, are reported in thesummary Table.

The compounds of the present invention which inhibit the binding of[³H]-bradykinin to the guinea-pig B₂ receptor (see Table I) also bind tothe human B₂ receptor cloned and transfected in a stable manner into CHOcells (Chinese Hamster Ovary cells). Thus, in this test, some compoundsinhibit the binding of [³H]-bradykinin to the B₂ receptor by at least95% at a concentration of 10 μM.

The compounds of the present invention can be useful in the treatment ofpain and particularly in the treatment of numerous pathologicalconditions involving bradykinin or its homologs. These pathologicalconditions include septic and hemorrhagic shock, anaphylactic reactions,arthrosis, rheumatoid polyarthritis, rhinitis, asthma, inflammatorydiseases of the gastrointestinal tract (for example colitis, rectitis,Crohn's disease), pancreatitis, certain carcinomas, hereditaryangioedema, migraine, encephalomyelitis, meningitis, cerebrovascularcomplications (especially those caused by cerebral traumatic shock),certain neurological disorders, vascular inflammatory states (forexample atherosclerosis and arteritis of the lower limbs), painfulstates (for example headache, toothache, menstrual pain), prematureuterine contractions, cystitis and burns. The compounds according to theinvention can also be useful for the potentiation of antiviral agents.

The compounds of the present invention, which can be used in the form ofthe free base or in the form of their non-toxic addition salts inassociation with a physiologically acceptable excipient, are generallyprescribed in human therapeutics at doses of about 1 to 1000 mg/day in aform which can be administered orally, by intravenous, intramuscular orsubcutaneous injection, transdermally, by means of aerosols or by meansof suppositories.

These compounds can also be administered topically, for example in theform of a gel or ointment.

The compounds of the present invention are also useful aspharmacological reagents, especially for the study of hormone-receptorinteractions. Use as a pharmacological reagent may require aradiolabeled derivative of one of the compounds according to theinvention (for example with tritium [³H] or sulfur [³⁵S]) in order toobtain a radioligand intended for conformational studies of thebradykinin B₂ receptor or for binding tests involving this type ofreceptor, for example for the evaluation of novel compounds which arecapable of exhibiting an affinity for the bradykinin B₂ receptor.

TABLE I

Activity Ex. Q R₁ A R₂ Salt % pK_(B)  1 Het 1 H —NH—(CH₂)₃—NH—CO— OH — 2 Het 1 H —NH—(CH₂)₃—NH—CO— H —  3 Het 1 H —NH—(CH₂)₃—NH—CO— H Ms  4Het 1 H

OH —  5 Het 1 H

H —  6 Het 1 H

H Ms  7 Het 1 CH₃ —NH—(CH₂)₃—NH—CO— OH —  8 Het 1 CH₃ —NH—(CH₂)₃—NH—CO—H —  9 Het 1 CH₃ —NH—(CH₂)₃—NH—CO— H Ms 97.4 8.6 10 Het 1 CH₃

OH — 11 Het 1 CH₃

H — 12 Het 1 CH₃

H Chl 13 Het 1 CH₃ —NH—CH₂— H — 14 Het 1 CH₃ —NH—CH₂— H Chl 15 Het 1 Br—NH—(CH₂)₃—NH—CO— H — 16 Het 1 Br —NH—(CH₂)₃—NH—CO— H Chl 100 9.1 17 Het1 Br

H Chl 100 8.4 18 Het 1 Br —NH—CH₂— H — 19 Het 1 Br —NH—CH₂— H Chl 20 Het2 — —NH—(CH₂)₃—NH—CO— OH — 21 Het 2 — —NH—(CH₂)₃—NH—CO— H 22 Het 2 ——NH—(CH₂)₃—NH—CO— H Ms 97.5 8.4 23 Het 2 —

OH 24 Het 2 —

H 25 Het 2 —

H Chl 26 Het 2 — —NH—CH₂— H — 27 Het 2 — —NH—CH₂— H Chl 28 Het 3 ——NH—(CH₂)₃—NH—CO— OH — 29 Het 3 — —NH—(CH₂)₃—NH—CO— H — 30 Het 3 ——NH—(CH₂)₃—NH—CO— H Ms 99.3 7.9 31 Het 3 —

OH — 32 Het 3 —

H — 33 Het 3 —

H Ms 96 7.8 34 Het 4 — —NH—CH₂— H — 35 Het 4 — —NH—CH₂— H Chl 36 Het 1Cl —NH—(CH₂)₃—NH—CO— OH — 37 Het 1 Cl —NH—(CH₂)₃—NH—CO— H — 38 Het 1 Cl—NH—(CH₂)₃—NH—CO— H Ms 100 39 Het 1 Cl

OH — 40 Het 1 Cl

H — 41 Het 1 Cl

H Ms 100 42 Het 1 —CO—CH₃ —NH—(CH₂)₃—NH—CO— OH — 43 Het 1 —CO—CH₃—NH—(CH₂)₃—NH—CO— H — 44 Het 1 —CO—CH₃ —NH—(CH₂)₃—NH—CO— H Ms 100 Notes:

Ms: addition salt with methanesulfonic acid Chl: addition salt withhydrochloric acid

What is claimed is:
 1. An[N-(Benzenesulfonyl)]N-(benzenesulfonyl)-L-proline compound selectedfrom the group consisting of: (i) the compounds of formula I:

 in which: X is a halogen atom, A is a divalent group —NH—(CH₂)_(n)—NH—CO—, —NH—CH₂— or

Q is a group

R₁ is a hydrogen atom, a halogen atom, a C₁-C₃ alkyl group with a linearor branched hydrocarbon chain, or a C₁-C₅ 1-oxoalkyl group, R₂ is ahydrogen atom or an OH group, and n is 2, 3 or 4; and (ii) theiraddition salts.
 2. The compound according to claim 1, wherein X is Cl.3. A therapeutic composition which contains, in association with aphysiologically acceptable excipient, at least one compound selectedfrom the group consisting of the compounds of formula I and theirnon-toxic addition salts according to claim
 1. 4. A method for thepreparation of a compound of formula I:

in which: X is a halogen atom, A is a divalent group—NH—(CH₂)_(n)—NH—CO—, —NH—CH₂—or

Q is a group selected from the group consisting of

R₁ is a hydrogen atom, a halogen atom, or a C₁-C₃ alkyl group with alinear or branched hydrocarbon chain, R₂ is H, or its addition salts,said method comprising: (1) reacting an alkali metal salt of ahydroxylated heterocyclic compound of the formula Q-O-Met  in which: Metis an alkali metal, and Q is a heterocyclic group selected from thegroup consisting of Het 1, Het 2, Het 3 and Het 4:

R_(1A) being a hydrogen atom or a C₁-C₃ alkyl group, with a compound ofthe formula II:

 in which X is a halogen atom and X₁ is a halogen atom, in an anhydroussolvent, at a temperature of between 0 and 50° C., for 0.5 to 10 hours,to give a compound of the formula III:

in which Q, X and R_(1A) are as defined above; (2) optionally, when Q inthe compound of formula III is Het 1 and R_(1A) is a hydrogen atom,reacting said compound of the formula III with a halogenating agent in asolvent, at a temperature of between about 0 and 50° C., for 0.5 to 20hours, to give a compound of the formula III′:

 in which: R_(1B) is a halogen atom, (3) hydrolyzing the ester group ofthe compound of the formula III or III′ at a temperature of from 20 to60° C., for 1 to 5 hours, to give a compound of the formula IV:

 in which Q and X are as defined above and R1 is a hydrogen atom, ahalogen atom or a C₁-C₃ alkyl group; and (4) reacting the compound ofthe formula IV with the salt of an amine of the formula

 wherein A is a group —NH—(CH₂)_(n)—NH—CO—, —NH—CH₂—or

in which n is 2, 3 or 4, in a solvent, at a temperature between 10-35°C., for 2 to 50 hours, to give the compound of the formula I.
 5. Themethod of claim 4, further comprising reacting the compound of theformula I, thus obtained, with an acid to give a corresponding acidaddition salt.
 6. A method for the preparation of a compound of theformula I:

in which: X is a halogen atom, A is a divalent group —NH—(CH₂)_(n)—NH—CO—, —NH—CH₂—or

Q is a group selected from the group consisting of

R₁ is a hydrogen atom, a halogen atom, or a C₁-C₃ alkyl group with alinear or branched hydrocarbon chain, and R₂ is OH or H, or its additionsalts, said method comprising: (1) reacting an alkali metal salt of ahydroxylated heterocyclic compound of the formula Q-O-Met  in which: Metis an alkali metal, and Q is a heterocyclic group selected from thegroup consisting of Het 1, Het 2, Het 3 and Het 4:

R_(1A) being a hydrogen atom or a C₁-C₃ alkyl group, with a compound ofthe formula II:

 in which X is a halogen atom and X₁ is a halogen atom, in an anhydroussolvent, at a temperature of between 0 and 50° C., for 0.5 to 10 hours,to give a compound of the formula III:

in which Q, X and R_(1A) are as defined above; (2) optionally, when Q inthe compound of formula III is Het 1 and R_(1A) is a hydrogen atom,reacting said compound of the formula III with a halogenating agent in asolvent, at a temperature of between 0 and 50° C., for 0.5 to 20 hours,to give a compound of the formula III′:

 in which R_(1B) is a halogen atom, (3) hydrolyzing the ester group ofthe compound of the formula III or III′ at a temperature of from 20 to60° C., for 1 to 5 hours, to give a compound of the formula IV:

 in which Q and X are as defined above and R₁ is a hydrogen atom, ahalogen atom or a C₁-C₃ alkyl group; (4) reacting the acid compound ofthe formula IV in which R₁ is a hydrogen atom, a chlorine atom or aC₁-C₃ alkyl group with a linear or branched hydrocarbon chain, with acompound of the formula VI:

 in which A is a group —NH—(CH₂)_(n)—NH—CO—, —NH—CH₂—or

in which n is 2, 3 or 4, in a solvent, at a temperature between 10-35°C., for 2 to 50 hours, to give a compound of the formula VII:

 in which Q, R₁, X and A are as defined above; (5) reacting the compoundof the formula VII with hydroxylamine in a solvent, at a temperaturebetween 15-25° C., for 1 to 12 hours, to give a compound of the formulaVIII:

 in which Q, R₁ , X and A are as defined above, said compound VIII beinga compound of the formula I wherein R₂ is OH; (6) acetylating thecompound of the formula VIII at a temperature between 10-35° C., for 1to 8 hours, to give a compound of the formula IX:

 in which Q, R₁, X and A are as defined above; and (7) reducing thecompound of the formula IV, thus obtained, by catalytic hydrogenation,at a temperature between 10-35° C., under a hydrogen pressure of between10⁵ and 10⁶ Pascals, to give the compound of the formula I in which Q,R₁, X and A are as defined above and R₂ is H.
 7. A method for thepreparation of a compound of the formula I:

in which X is a halogen atom, A is a divalent group  —NH—(CH₂)₂—NH—CO—,—NH—CH₂— or

Q is a group selected from the group consisting of

R₁ is a hydrogen atom, a halogen atom, or a C₁-C₃ alkyl group with alinear or branched hydrocarbon chain, R₂ is H, or its addition salts,said method comprising: (1) reacting an alkali metal salt of ahydroxylated heterocyclic compound of the formula Q-O-Met  in which: Metis an alkali metal, and Q is a heterocyclic group selected from thegroup consisting of Het 1, Het 2, Het 3 and Het 4:

R_(1A) being a hydrogen atom or a C₁-C₃ alkyl group, with a compound ofthe formula II:

 in which X is a halogen atom and X₁ is a halogen atom, in an anhydroussolvent, at a temperature of between 0 and 50° C., for 0.5 to 10 hours,to give a compound of the formula III:

in which Q, X and R_(1A) are as defined above; (2) optionally, when Q inthe compound of formula III is Het 1 in which R_(1A) is a hydrogen atom,reacting said compound of the formula III with a halogenating agent in asolvent, at a temperature of between 0 and 50° C., for 0.5 to 20 hours,to give a compound of the formula III′:

 in which: R_(1B) is a halogen atom, (3) hydrolyzing the ester group ofthe compound of the formula III or III′ at a temperature of from 20 to60° C., for 1 to 5 hours, to give a compound of the formula IV:

 in which Q and X are as defined above and R₁ is a hydrogen atom, ahalogen atom or a C₁-C₃ alkyl group; (4) reacting the compound of theformula IV with an amine of the formula H₂N—(CH₂)_(n)—NH—R₃ or

 in which n is 2, 3 or 4 and R₃ is an amino-protecting group, in asolvent, at a temperature between 10-35° C., for 2 to 50 hours, to givea compound of the formula X:

in which B is —NH—(CH2)_(n)—NH— or

and Q, R₁, R₃ n and X are as defined above; (5) deprotecting the aminegroup of the compound of the formula X so as to replace the group R₃with a hydrogen atom, to give a compound of the formula XI:

 in which B, Q, R₁ and X are as defined above; and 6) reacting thecompound of the formula XI with 4-(aminoiminomethyl)benzoic acid, in asolvent, at a temperature between 10-35° C., for 2 to 50 hours, to givea compound of the formula I:

 in which: Q, R₁ and X are as defined above, R₂ is H, A is a group

 and n is as defined above.
 8. A method for the preparation of acompound of the formula I:

in which: X is a halogen atom, A is a divalent group—NH—(CH₂)_(n)—NH—CO—, —NH—CH₂— or

n is 2, 3 or 4, Q is a group selected from the group consisting of

R₁ is a hydrogen atom, a halogen atom, a C₁-C₃ alkyl group with a linearor branched hydrocarbon chain, or a C₁-C₅ 1-oxoalkyl group, and R₂ is OHor H, or its addition salts, said method comprising: (1) reacting anacid of the formula XII:

 in which X is a halogen atom, with a compound of the formula

in which A is a group —NH—(CH₂)_(n)—NH—CO—, —NH—CH₂— or

in which n is 2, 3 or 4, in a solvent, at a temperature between 10-35°C., for 2 to 50 hours, to give a compound of the formula XIII:

 in which A and X are as defined above; (2) reacting the compound of theformula XIII with a hydroxylated heterocyclic compound of the formulaQ-OH, Q being selected from the group consisting of the structures:

 in which R₁ is a hydrogen atom, a halogen atom, a C₁-C₃ alkyl group ora C₁-C₅ 1-oxoalkyl group, in an anhydrous solvent, at a temperature ofbetween 0 and 50° C., for 0.5 to 10 hours, to give a compound of theformula XIV:

in which Q, R₁, X and A are as defined above; (3) reacting the compoundof the formula XIV with hydroxylamine in a solvent, at between 15-25°C., for 1 to 12 hours, to give a compound of the formula VIII:

 in which Q, R₁, X and A are as defined above, said compound VIII beinga compound of the formula I wherein R₂ is OH; (4) acetylating thecompound of the formula VIII at a temperature between 10-35° C., for 1to 8 hours, to give a compound of the formula IX:

 in which Q, R₁, X and A are as defined above; and (5) reducing thecompound of the formula IX by catalytic hydrogenation, at a temperaturebetween 10-35° C., under a hydrogen pressure of between 10⁵ and 10⁶Pascals, to give a compound of the formula I in which Q, R₁, X and A areas defined above and R₂ is H.